Recombinant Anti-LLGL1 + LLGL2 antibody [EPR18899] (ab183021)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18899] to LLGL1 + LLGL2
- Suitable for: IHC-P, WB, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-LLGL1 + LLGL2 antibody [EPR18899]
See all LLGL1 + LLGL2 primary antibodies -
Description
Rabbit monoclonal [EPR18899] to LLGL1 + LLGL2 -
Host species
Rabbit -
Specificity
The immunogen used for this product shares 53% identity (67% positives) with LLGL2. Cross-reactivity with this protein has not been confirmed experimentally. -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, SW480, A431, Caco-2 and 293 whole cell lysates; Human fetal liver, fetal heart and fetal kidney lysates. IHC-P: Human kidney and stomach tissues. ICC/IF: HepG2 and SW480 cells. IP: HepG2 whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18899 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab183021 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/2000. Detects a band of approximately 115 kDa (predicted molecular weight: 115 kDa).
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ICC/IF |
1/100.
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IP |
1/50.
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Notes |
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/2000. Detects a band of approximately 115 kDa (predicted molecular weight: 115 kDa). |
ICC/IF
1/100. |
IP
1/50. |
Target
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Relevance
LLGL1 is a protein that is similar to a tumor suppressor in Drosophila. The protein is part of a cytoskeletal network and is associated with nonmuscle myosin II heavy chain and a kinase that specifically phosphorylates this protein at serine residues. The gene for LLGL1 is located within the Smith-Magenis syndrome region on chromosome 17. LLGL2 is a protein similar to lethal (2) giant larvae of Drosophila. In fly, the protein's ability to localize cell fate determinants is regulated by the atypical protein kinase C (aPKC). In human, this protein interacts with aPKC-containing complexes and is cortically localized in mitotic cells. -
Cellular localization
Cytoplasmic -
Database links
- Entrez Gene: 3993 Human
- Entrez Gene: 3996 Human
- Omim: 600966 Human
- SwissProt: Q15334 Human
- SwissProt: Q6P1M3 Human
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Alternative names
- DLG4 antibody
- HGL antibody
- HUGL antibody
see all
Images
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All lanes : Anti-LLGL1 + LLGL2 antibody [EPR18899] (ab183021) at 1/2000 dilution
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method
Lane 3 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method
Lane 4 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method
Lane 5 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method
Lane 6 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 115 kDa
Observed band size: 115 kDa
Exposure time: 20 secondsBlocking buffer: 5% NFDM/TBST.
We recommend to use 1% SDS Hot lysis method to get desired WB results.
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All lanes : Anti-LLGL1 + LLGL2 antibody [EPR18899] (ab183021) at 1/10000 dilution
Lane 1 : His tagged human LLGL1 full length recombinant protein
Lane 2 : Flag tagged human LLGL2 full length recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 115 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Exposure time: 40 secondsBlocking buffer: 5% NFDM/TBST.
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All lanes : Anti-LLGL1 + LLGL2 antibody [EPR18899] (ab183021) at 1/5000 dilution
Lane 1 : HepG2 cell lysate
Lane 2 : A431 cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : A459 cell lysate
Lane 5 : Caco-2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 115 kDa
Observed band size: 115 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : Anti-LLGL1 + LLGL2 antibody [EPR18899] (ab183021) at 1/2000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 115 kDa
Observed band size: 115 kDaBlocking/dilution buffer: 5% NFDM/TBST.
Exposure time Lane 1/2: 3 minutes; Lane 3: 15 seconds.
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling LLGL1 with ab183021 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human kidney is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling LLGL1 with ab183021 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human stomach is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling LLGL1 with ab183021 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [EPR18899]-Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab183021 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (Human colorectal adenocarcinoma cell line) cells labeling LLGL1 with ab183021 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SW480 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [EPR18899]-Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab183021 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution. -
LLGL1was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab183021 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab183021 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 whole cell lysate 10µg (Input).
Lane 2: ab183021 IP in HepG2 whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR18899] - Isotype Control (ab172730) instead of ab183021 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab183021 has been referenced in 1 publication.
- Tocan V et al. Hepatocyte polarity establishment and apical lumen formation are organized by Par3, Cdc42, and aPKC in conjunction with Lgl. J Biol Chem 297:101354 (2021). PubMed: 34717957